TABLE 2.
Bacterial strains and plasmids used in the study
| Strain or plasmid | Relevant characteristicsa | Reference or origin |
|---|---|---|
| Strain | ||
| Nocardioides sp. PD653 | HCB+, PCP+ | 13 |
| Nocardioides sp. PD653-B2 | HCB−, PCP+ | This study |
| E. coli DH5α | F−, l−, f80dlacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rK−, mK+), pho A, supE44, thi-1, gyrA96, relA1 | Toyobo |
| E. coli BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm λ(DE3); T7 RNA polymerase gene under the control of the lacUV promoter | Novagen |
| Plasmid | ||
| pGEM-T Easy | Apr lacZ, pMB1-derived replicon, TA cloning vector | Promega |
| pGEM-T Easy::2177-2179 | Apr, pGEM-T Easy with 2.9-kb PCR-amplified DNA fragment containing ORF1, ORF2, and ORF3 of PD653 | This study |
| pETDuet-1 | Apr pBR322-derived ColE1 replication, T7 promoter, two MCS, expression vector | Novagen |
| pE123N | Apr, pE12N with 0.7-kb PCR-amplified DNA fragment containing ORF3 in MCS2 | This study |
| pE12N | Apr, pETDuet-1with PCR-amplified DNA fragment containing ORF1 and ORF2 in MCS1 | This study |
| pE12N2 | Apr, pE1N with PCR-amplified DNA fragment containing ORF2 in MCS2 | This study |
| pE23N2 | Apr, pE2N with PCR-amplified DNA fragment containing ORF3 in MCS2 | This study |
| pE13N2 | Apr, pE1N with PCR-amplified DNA fragment of ORF3 in MCS2 | This study |
| pE1N | Apr, pETDuet-1 with PCR-amplified DNA fragment of ORF1 in MCS1 | This study |
| pE2N | Apr, pETDuet-1 with PCR-amplified DNA fragment of ORF2 in MCS1 | This study |
| pE3N | Apr, pETDuet-1 with PCR-amplified DNA fragment of ORF3 in MCS1 | This study |
a
HCB+, able to degrade hexachlorobenzene (HCB); HCB−, unable to degrade HCB; PCP+, able to degrade pentachlorophenol (PCP); PCP−, unable to degrade PCP; Apr, ampicillin resistant.